Page 8 - 南京医科大学自然版
P. 8

第45卷第1期
               · 2  ·                            南 京    医 科 大 学 学         报                        2025年1月


              divided into a negative control（NC）group，a Sham group，a CLP group，a CLP + MON group，a Sham + MON group，and a CLP +
              dexamethasone（DEX）group. Drug or equivalent saline was injected intraperitoneally once daily for consecutive five days after CLP.
              After all mice were euthanized on day 5，serum and kidney tissues were collected for subsequent experiments. Blood urea nitrogen
             （BUN）and creatinine（CRE）concentrations in the serum were detected by biochemical kits，as well as renal oxidative stress related
              indicators，such as glutathione（GSH），catalase（CAT），total antioxidant capacity（T⁃AOC）and malondialdehyde（MDA）levels. The
              pathological changes of renal tissues were observed by the H&E staining，and the levels of reactive oxygen species（ROS）in renal
              tissues stained with dihydroethidium（DHE）were observed by laser confocal microscopy. The levels of tumor necrosis factor⁃α（TNF⁃
              α），interleukin（IL）⁃1β and IL⁃6 in the serum and kidney tissues were detected by ELISA and RT⁃qPCR. Western blot was used to
              detect the protein expression levels of NLR family pyrin domain containing protein 3（NLRP3）inflammasome and nuclear factor kappa⁃
              B（NF⁃κB）signaling pathway. In addition，lipopolysaccharide（LPS）/adenosine triphosphate（ATP）was used to induce HK⁃2 cells to
              establish an in vitro sepsis model. Cell experiments were divided into a NC group，a LPS/ATP group，a LPS/ATP + MON group，a
                   OE                           OE
              NLRP3 + LPS/ATP+MON group and a IKKβ + LPS/ATP+MON group. CCK⁃8 was used to detect the cell viability，and ELISA was
              used to detect the secretion of inflammatory cytokines in HK⁃2 cells. Results：Compared with the CLP group，the survival rate of mice
              in the CLP+MON group was significantly increased，the appearance of renal tissues was restored from black to bright red，the serum
              BUN and CRE levels were significantly decreased，and the abnormal pathological changes of renal tissues with increased inflammatory
              cells were significantly improved. Compared with the CLP group，the levels of TNF⁃α，IL⁃1β，and IL⁃6 were significantly decreased，
              GSH，CAT，and T⁃AOC levels were increased，and MDA and ROS levels were significantly decreased in the MON treatment group.
              Western blot results showed that compared with the CLP group，the expression levels of NLRP3，Caspase⁃1，Cleaved⁃Caspase⁃1，and
              p⁃NF⁃κB P65 protein in the CLP+MON group decreased significantly，but the expression levels of ihibitor of nuclear factor kappa⁃B α
                                                    OE
                                                                               OE
             （IκBα）increased significantly. In addition，NLRP3 +LPS/ATP+MON group and IKKβ +LPS/ATP+MON group activated NLRP3
              inflammasome and NF⁃κB pathway，and reversed the inhibitory effect of MON on inflammatory cytokines in the LPS/ATP⁃stimulated
              HK⁃2 cells，compared with the LPS/ATP+MON group. Conclusion：MON reduces the release of inflammatory factors by inhibiting the
              NF⁃κB/NLRP3 inflammasome pathway to improve mouse S⁃AKI and dysfunction.
             ［Key words］ monotropein；sepsis⁃associated acute kidney injury；NLRP3 inflammasome；NF⁃κB
                                                                               ［J Nanjing Med Univ，2025，45（01）：1⁃12］




                  脓毒症是由细菌、真菌、病毒及寄生虫等引起                          键因素 。S⁃AKI 的主要原因是病原相关分子模式
                                                                      ［4］
              的全身炎症反应综合征，常见于严重的创伤或感染                           （pathogen ⁃ associated molecular patterns，PAMP）和
              性疾病患者。器官衰竭在脓毒症病理生理过程中                             损伤相关分子模式（damage⁃associated molecular pat⁃
              起着至关重要的作用，对脓毒症的死亡率有着直接                            terns，DAMP）的持续刺激。脓毒症发生时，PAMP和
                          ［1］
              且显著的影响 。肾脏是脓毒症中最先受损的器官                            DAMP激活免疫系统，启动细胞内的分子级联反应，
              之一，高达2/3的脓毒症或脓毒性休克患者会发生急                          进而引发炎症反应和氧化应激，最终导致肾组织损
                                                                                 ［5］
              性肾损伤。脓毒症相关急性肾损伤（sepsis⁃associated                 伤和器官功能障碍 。
              acute kidney injury，S⁃AKI）是脓毒症的重要并发症之                  核因子⁃κB（nuclear factor kappa⁃B，NF⁃κB）是一
              一，具有较高的发病率和死亡率，已成为全球重要的                           种转录因子，NF⁃κB 通路在脓毒症的发病机制中起
                          ［2］
              公共健康问题 。了解S⁃AKI的发病机制将为其早期                         着关键作用，它能够调控多种炎症相关基因的表
              诊断和治疗策略的发展奠定基础，对于提高脓毒症患                           达，例如促炎细胞因子肿瘤坏死因子（tumor necrosis
              者的生存率和生活质量具有重要意义。                                 factor，TNF）⁃α和白介素（interleukin，IL）⁃1β。抑制NF⁃
                  S⁃AKI 的发病机制非常复杂，涉及多种因素的                       κB通路可以阻断过度的炎症反应，从而减轻组织损
                                                                  ［6］
              协同作用，主要包括异常的炎症调节、氧化应激和                            伤 。研究表明，NF⁃κB通路的激活可以调控NLR家
                        ［3］
              细胞凋亡等 。炎症反应是宿主抵抗感染的主要防                            族Pyrin域蛋白3（NLR family pyrin domain containing
              御机制，在启动和介导损伤后功能恢复的过程中发                            protein 3，NLRP3）炎症小体的表达和激活。此外，活
              挥着至关重要的作用。当机体遭遇有害刺激时，正                            性氧（reactive oxygen species，ROS）也可以触发NF⁃κB

              常的炎症反应会被触发以进行防御。然而在S⁃AKI                          介导的NLRP3炎症小体激活。NLRP3炎性小体是由
              中，这种炎症反应会出现失调，主要表现为过度激                            NLRP3、含CARD的凋亡相关斑点样蛋白（apoptosis⁃
              活。炎症反应失调是导致S⁃AKI发病过程加剧的关                          associated speck ⁃ like protein containing a CARD，
   3   4   5   6   7   8   9   10   11   12   13